RESUMEN
OBJECTIVES: Specialised pro-resolving mediator (SPM) can dampen the acute inflammation through ERV1, ALX/FPR2 and BLT1 cell receptors and it is conceivable that their expression is dysregulated during chronic inflammation. The aim of this study was to evaluate the expression of ERV1, ALX/FPR2 and BLT1 on peripheral blood (PB) cells from rheumatoid arthritis (RA) patients. METHODS: At baseline, erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), clinimetric indexes (28-joint disease activity score (DAS28) and clinical disease activity index (CDAI)), and PB samples were collected from 33 RA patients. Based on DAS28, patients were divided into high-moderate (H-Mo/RA, DAS28≥3.2) and low-remission (L-Rem/RA, DAS28<3.2) disease activity group. Cell membrane expression of ERV1, ALX/FPR2 and BLT1 on CD3pos, CD19pos, CD14pos cells and granulocytes was assessed by multi-parametric flow-cytometry analysis. Nine healthy controls (HC) were also studied. RESULTS: Sixteen H-Mo/RA and 17 L-Rem/RA patients were identified. The percentage of BLT1posCD14pos cells was significantly higher in L-Rem/RA (47.17%) than in H-Mo/RA (14.27%) group (p=0.005). Likewise, the percentage ALX/FPR2pos CD14pos cells was significantly higher in L-Rem/RA (33.02%) than in H-Mo/RA (8.77%; p=0.04) patients. An inverse correlation between BLT1posCD14pos cell percentage and DAS28 (r=-0.42; p=0.01), CDAI (r=-0.51; p=0.003), ESR (r=-0.39; p=0.025) and CRP (r=-0.40; p=0.02), ALX/FPR2posCD14pos cell percentage and CRP (r=-0.39; p=0.02) were found, while SPM-receptors mean fluorescence intensity (MFI) was not different between HC and L-Rem/RA patients. CONCLUSIONS: ALX/FPR2 and BLT1 receptors expression mirrors RA disease activity arising as potential biomarkers of inflammatory regulation.
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Artritis Reumatoide , Monocitos , Proteínas Adaptadoras Transductoras de Señales/inmunología , Artritis Reumatoide/diagnóstico , Biomarcadores , Sedimentación Sanguínea , Proteína C-Reactiva/análisis , Humanos , Inflamación , Receptores de Lipopolisacáridos/inmunología , Receptores de Formil Péptido , Receptores de Leucotrieno B4/inmunología , Receptores de Lipoxina , Índice de Severidad de la EnfermedadRESUMEN
Retinal vascular diseases have distinct, complex and multifactorial pathogeneses yet share several key pathophysiological aspects including inflammation, vascular permeability and neovascularisation. In non-infectious posterior uveitis (NIU), retinal vasculitis involves vessel leakage leading to retinal enlargement, exudation, and macular oedema. Neovascularisation is not a common feature in NIU, however, detection of the major angiogenic factor-vascular endothelial growth factor A (VEGF-A)-in intraocular fluids in animal models of uveitis may be an indication for a role for this cytokine in a highly inflammatory condition. Suppression of VEGF-A by directly targeting the leukotriene B4 (LTB4) receptor (BLT1) pathway indicates a connection between leukotrienes (LTs), which have prominent roles in initiating and propagating inflammatory responses, and VEGF-A in retinal inflammatory diseases. Further research is needed to understand how LTs interact with intraocular cytokines in retinal inflammatory diseases to guide the development of novel therapeutic approaches targeting both inflammatory mediator pathways.
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Inflamación/tratamiento farmacológico , Receptores de Leucotrieno B4/metabolismo , Vasculitis Retiniana/metabolismo , Factor A de Crecimiento Endotelial Vascular/metabolismo , Animales , Modelos Animales de Enfermedad , Humanos , Receptores de Leucotrieno B4/inmunología , Vasculitis Retiniana/inmunología , Uveítis/tratamiento farmacológico , Uveítis/inmunologíaRESUMEN
Increased levels of neutrophils in bronchoalveolar lavage fluid (BALF) were associated with asthma severity. As leukotriene B4 (LTB4) is a principal chemoattractant molecule for neutrophils, its receptors, BLT1 and BLT2, may contribute to neutrophil-dominant airway inflammation. In the present study, we established a mouse model of steroid-resistant, neutrophil-dominant airway inflammation by house dust mite (HDM)/lipopolysaccharide (LPS) sensitization and HDM challenge, and we investigated whether BLT1/BLT2 signaling was associated with the development of neutrophilic airway inflammation. Blockade of BLT1 or BLT2 significantly suppressed airway inflammation and IL-17 production in this mouse model. The 5-LO and 12-LO enzymes, which catalyze the synthesis of BLT1/BLT2 ligands, were also critically associated with neutrophil-dominant airway inflammation and the synthesis of IL-17. Collectively, our results suggest that the 5-/12-LO-BLT1/BLT2-linked cascade significantly contributes to neutrophil-dominant severe airway inflammation via IL-17 synthesis in HDM-induced neutrophilic asthma.
Asunto(s)
Asma/etiología , Interleucina-17/biosíntesis , Neutrófilos/inmunología , Pyroglyphidae/inmunología , Receptores de Leucotrieno B4/inmunología , Alérgenos/inmunología , Animales , Araquidonato 12-Lipooxigenasa/metabolismo , Araquidonato 5-Lipooxigenasa/metabolismo , Asma/inmunología , Asma/metabolismo , Modelos Animales de Enfermedad , Femenino , Inflamación/etiología , Inflamación/inmunología , Inflamación/metabolismo , Pulmón/inmunología , Pulmón/metabolismo , Pulmón/patología , Ratones , Ratones Endogámicos BALB C , Neutrófilos/patología , Transducción de Señal/inmunologíaRESUMEN
We investigated the effects of resolvin E (RvE) 1, RvE2, and RvE3 on IL-4- and IL-33-stimulated bone marrow-derived dendritic cells (BMDCs) from house dust mite (HDM)-sensitized mice. We also investigated the role of RvE3 in a murine model of HDM-induced airway inflammation. In vitro, BMDCs from HDM-sensitized mice were stimulated with IL-4 and IL-33 and then treated with RvE1, RvE2, RvE3, or vehicle. RvE1, RvE2, and RvE3 suppressed IL-23 release from BMDCs. In vivo, RvE3 administrated to HDM-sensitized and challenged mice in the resolution phase promoted a decline in total numbers of inflammatory cells and eosinophils, reduced levels of IL-23 and IL-17 in lavage fluid, and suppressed IL-23 and IL-17A mRNA expression in lung and peribronchial lymph nodes. RvE3 also reduced resistance in the lungs of HDM-sensitized mice. A NanoBiT ß-arrestin recruitment assay using human embryonic kidney 293 cells revealed that pretreatment with RvE3 suppressed the leukotriene B4 (LTB4)-induced ß-arrestin 2 binding to LTB4 receptor 1 (BLT1R), indicating that RvE3 antagonistically interacts with BLT1R. Collectively, these findings indicate that RvE3 facilitates the resolution of allergic airway inflammation, partly by regulating BLT1R activity and selective cytokine release by dendritic cells. Our results accordingly identify RvE3 as a potential therapeutic target for the management of asthma.-Sato, M., Aoki-Saito, H., Fukuda, H., Ikeda, H., Koga, Y., Yatomi, M., Tsurumaki, H., Maeno, T., Saito, T., Nakakura, T., Mori, T., Yanagawa, M., Abe, M., Sako, Y., Dobashi, K., Ishizuka, T., Yamada, M., Shuto, S., Hisada, T. Resolvin E3 attenuates allergic airway inflammation via the interleukin-23-interleukin-17A pathway.
Asunto(s)
Asma/inmunología , Ácidos Grasos Insaturados/inmunología , Interleucina-17/inmunología , Subunidad p19 de la Interleucina-23/inmunología , Transducción de Señal/inmunología , Animales , Asma/patología , Células de la Médula Ósea/inmunología , Células de la Médula Ósea/patología , Células Dendríticas/inmunología , Células Dendríticas/patología , Femenino , Regulación de la Expresión Génica/inmunología , Leucotrieno B4/inmunología , Ratones , Ratones Endogámicos BALB C , Pyroglyphidae/inmunología , Receptores de Leucotrieno B4/inmunología , Arrestina beta 2/inmunologíaRESUMEN
Leukotriene B4 (LTB4 ) plays a prominent role in innate immunity as it induces phagocyte recruitment, the release of antimicrobial effectors, and as it potentiates the ingestion and killing of pathogens. In humans, LTB4 has a short half-life and is rapidly metabolized by leukocytes, notably into 20-OH- and 20-COOH-LTB4 by neutrophils. Although these LTB4 metabolites bind to the BLT1 receptor with high affinity, they activate neutrophils to a much lower extent than LTB4 . We thus postulated that LTB4 metabolites could dampen BLT1 -mediated responses, therefore limiting the impact of LTB4 on human neutrophil functions. We found that 20-OH-LTB4 and 20-COOH-LTB4 inhibited all of the LTB4 -mediated neutrophil responses we tested (migration, degranulation, leukotriene biosynthesis). The potencies of the different compounds at inhibiting LTB4 -mediated responses were 20-OH-LTB4 = CP 105,696 (BLT1 antagonist) > > 20-COOH-LTB4 ≥ resolvin E1 (RVE1 ). In contrast, the fMLP- and IL-8-mediated responses we tested were not affected by the LTB4 metabolites or RVE1 . 20-OH-LTB4 and 20-COOH-LTB4 also inhibited the LTB4 -mediated migration of human eosinophils but not that induced by 5-KETE. Moreover, using 20-COOH-LTB4 , LTB4 , and LTB4 -alkyne, we show that LTB4 is a chemotactic, rather than a chemokinetic factor for both human neutrophils and eosinophils. In conclusion, our data indicate that LTB4 metabolites and RVE1 act as natural inhibitors of LTB4 -mediated responses. Thus, preventing LTB4 ω-oxidation might result in increased innate immunity and granulocyte functions.
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Eosinófilos/inmunología , Leucotrieno B4/inmunología , Neutrófilos/inmunología , Receptores de Leucotrieno B4/inmunología , Ácidos Araquidónicos/farmacología , Benzopiranos/farmacología , Ácidos Carboxílicos/farmacología , Ácido Eicosapentaenoico/análogos & derivados , Ácido Eicosapentaenoico/farmacología , Eosinófilos/citología , Humanos , Leucotrieno B4/farmacología , Neutrófilos/citología , Receptores de Leucotrieno B4/antagonistas & inhibidoresRESUMEN
Given the importance of high-mobility group box 1 (HMGB1) and 5-lipoxygenase (5-LO) signaling in vascular inflammation, we investigated the role of leukotriene signaling in monocytes on monocyte-to-macrophage differentiation (MMD) induced by HMGB1, and on vascular inflammation and subsequent intimal hyperplasia in a mouse model of wire-injured femoral artery. In cultured primary bone marrow-derived cells (BMDCs) stimulated with HMGB1, the number of cells with macrophage-like morphology was markedly increased in association with an increased expression of CD11b/Mac-1, which were attenuated in cells pre-treated with Zileuton, a 5-LO inhibitor as well as in 5-LO-deficient BMDCs. Of various leukotriene receptor inhibitors examined, which included leukotriene B4 receptors (BLTRs) and cysteinyl leukotriene receptors (cysLTRs), the BLTR1 inhibitor (U75302) exclusively suppressed MMD induction by HMGB1. The importance of BLTR1 in HMGB1-induced MMD was also observed in BMDCs isolated from BLTR1-deficient mice and BMDCs transfected with BLTR1 siRNA. Although leukotriene B4 (LTB4) had minimal direct effects on MMD in control and 5-LO-deficient BMDCs, MMD attenuation by HMGB1 in 5-LO-deficient BMDCs was significantly reversed by exogenous LTB4, but not in BLTR1-deficient BMDCs, suggesting that LTB4/BLTR1-mediated priming of monocytes is a prerequisite of HMGB1-induced MMD. In vivo, both macrophage infiltration and intimal hyperplasia in our wire-injured femoral artery were markedly attenuated in BLTR1-deficient mice as compared with wild-type controls, but these effects were reversed in BLTR1-deficient mice transplanted with monocytes from control mice. These results suggest that BLTR1 in monocytes is a pivotal player in MMD with subsequent macrophage infiltration into neointima, leading to vascular remodeling after vascular injury.
Asunto(s)
Alcoholes Grasos/farmacología , Arteria Femoral , Glicoles/farmacología , Monocitos , Receptores de Leucotrieno B4 , Remodelación Vascular , Lesiones del Sistema Vascular , Vasculitis , Animales , Arteria Femoral/inmunología , Arteria Femoral/lesiones , Arteria Femoral/patología , Hiperplasia , Macrófagos/inmunología , Macrófagos/patología , Ratones , Ratones Noqueados , Monocitos/inmunología , Monocitos/patología , Receptores de Leucotrieno B4/antagonistas & inhibidores , Receptores de Leucotrieno B4/genética , Receptores de Leucotrieno B4/inmunología , Remodelación Vascular/efectos de los fármacos , Remodelación Vascular/genética , Remodelación Vascular/inmunología , Lesiones del Sistema Vascular/tratamiento farmacológico , Lesiones del Sistema Vascular/genética , Lesiones del Sistema Vascular/inmunología , Lesiones del Sistema Vascular/patología , Vasculitis/tratamiento farmacológico , Vasculitis/genética , Vasculitis/inmunología , Vasculitis/patologíaRESUMEN
The presence of mast cells in some human colorectal cancers is a positive prognostic factor, but the basis for this association is incompletely understood. Here, we found that mice with a heterozygous mutation in the adenomatous polyposis coli gene (ApcMin/+) displayed reduced intestinal tumor burdens and increased survival in a chemokine decoy receptor, ACKR2-null background, which led to discovery of a critical role for mast cells in tumor defense. ACKR2-/-ApcMin/+ tumors showed increased infiltration of mast cells, their survival advantage was lost in mast cell-deficient ACKR2-/-SA-/-ApcMin/+ mice as the tumors grew rapidly, and adoptive transfer of mast cells restored control of tumor growth. Mast cells from ACKR2-/- mice showed elevated CCR2 and CCR5 expression and were also efficient in antigen presentation and activation of CD8+ T cells. Mast cell-derived leukotriene B4 (LTB4) was found to be required for CD8+ T lymphocyte recruitment, as mice lacking the LTB4 receptor (ACKR2-/-BLT1-/-ApcMin/+) were highly susceptible to intestinal tumor-induced mortality. Taken together, these data demonstrate that chemokine-mediated recruitment of mast cells is essential for initiating LTB4/BLT1-regulated CD8+ T-cell homing and generation of effective antitumor immunity against intestinal tumors. We speculate that the pathway reported here underlies the positive prognostic significance of mast cells in selected human tumors. Cancer Immunol Res; 6(3); 332-47. ©2018 AACR.
Asunto(s)
Linfocitos T CD8-positivos/inmunología , Neoplasias Intestinales/inmunología , Mastocitos/inmunología , Proteína de la Poliposis Adenomatosa del Colon/genética , Proteína de la Poliposis Adenomatosa del Colon/inmunología , Animales , Femenino , Vigilancia Inmunológica , Leucotrieno B4/inmunología , Masculino , Ratones Transgénicos , Receptores de Quimiocina/genética , Receptores de Quimiocina/inmunología , Receptores de Leucotrieno B4/genética , Receptores de Leucotrieno B4/inmunologíaRESUMEN
Leukotriene B4 (LTB4) receptor 1 (BLT1) is a G protein-coupled receptor expressed in various leukocyte subsets; however, the precise expression of mouse BLT1 (mBLT1) has not been reported because a mBLT1 monoclonal antibody (mAb) has not been available. In this study, we present the successful establishment of a hybridoma cell line (clone 7A8) that produces a high-affinity mAb for mBLT1 by direct immunization of BLT1-deficient mice with mBLT1-overexpressing cells. The specificity of clone 7A8 was confirmed using mBLT1-overexpressing cells and mouse peripheral blood leukocytes that endogenously express BLT1. Clone 7A8 did not cross-react with human BLT1 or other G protein-coupled receptors, including human chemokine (C-X-C motif) receptor 4. The 7A8 mAb binds to the second extracellular loop of mBLT1 and did not affect LTB4 binding or intracellular calcium mobilization by LTB4. The 7A8 mAb positively stained Gr-1-positive granulocytes, CD11b-positive granulocytes/monocytes, F4/80-positive monocytes, CCR2-high and CCR2-low monocyte subsets in the peripheral blood and a CD4-positive T cell subset, Th1 cells differentiated in vitro from naïve CD4-positive T cells. This mAb was able to detect Gr-1-positive granulocytes and monocytes in the spleens of naïve mice by immunohistochemistry. Finally, intraperitoneal administration of 7A8 mAb depleted granulocytes and monocytes in the peripheral blood. We have therefore succeeded in generating a high-affinity anti-mBLT1 mAb that is useful for analyzing mBLT1 expression in vitro and in vivo.
Asunto(s)
Anticuerpos Monoclonales de Origen Murino/inmunología , Leucotrieno B4/inmunología , Receptores de Leucotrieno B4/antagonistas & inhibidores , Animales , Anticuerpos Monoclonales de Origen Murino/química , Anticuerpos Monoclonales de Origen Murino/farmacología , Células CHO , Señalización del Calcio/efectos de los fármacos , Diferenciación Celular/inmunología , Cricetinae , Cricetulus , Granulocitos/inmunología , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Noqueados , Monocitos/inmunología , Estructura Secundaria de Proteína , Receptores de Leucotrieno B4/química , Receptores de Leucotrieno B4/inmunología , Células TH1/inmunologíaRESUMEN
Tissue inflammation is a key component of obesity-induced insulin resistance, with a variety of immune cell types accumulating in adipose tissue. Here, we have demonstrated increased numbers of B2 lymphocytes in obese adipose tissue and have shown that high-fat diet-induced (HFD-induced) insulin resistance is mitigated in B cell-deficient (Bnull) mice. Adoptive transfer of adipose tissue B2 cells (ATB2) from wild-type HFD donor mice into HFD Bnull recipients completely restored the effect of HFD to induce insulin resistance. Recruitment and activation of ATB2 cells was mediated by signaling through the chemokine leukotriene B4 (LTB4) and its receptor LTB4R1. Furthermore, the adverse effects of ATB2 cells on glucose homeostasis were partially dependent upon T cells and macrophages. These results demonstrate the importance of ATB2 cells in obesity-induced insulin resistance and suggest that inhibition of the LTB4/LTB4R1 axis might be a useful approach for developing insulin-sensitizing therapeutics.
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Tejido Adiposo/inmunología , Subgrupos de Linfocitos B/inmunología , Resistencia a la Insulina/inmunología , Leucotrieno B4/inmunología , Receptores de Leucotrieno B4/inmunología , Transducción de Señal/inmunología , Tejido Adiposo/patología , Animales , Subgrupos de Linfocitos B/patología , Grasas de la Dieta/efectos adversos , Grasas de la Dieta/farmacología , Resistencia a la Insulina/genética , Leucotrieno B4/genética , Macrófagos/inmunología , Macrófagos/patología , Ratones , Ratones Noqueados , Obesidad/inducido químicamente , Obesidad/genética , Obesidad/inmunología , Obesidad/patología , Receptores de Leucotrieno B4/genética , Transducción de Señal/genética , Linfocitos T/inmunología , Linfocitos T/patologíaRESUMEN
Unresolved inflammation is key in linking metabolic dysregulation and the immune system in type 2 diabetes. Successful regulation of acute inflammation requires biosynthesis of specialized proresolving lipid mediators, such as E-series resolvin (RvE) 1, and activation of cognate G protein-coupled receptors. RvE1 binds to leukotriene B4 (BLT-1) on neutrophils and to ERV-1/ChemR23 on monocyte/macrophages. We show novel actions of RvE1 and expression patterns of neutrophil receptors in type 2 diabetes. Neutrophils from healthy subjects express functional BLT-1, low levels of minimally functional ERV-1, and inversed coexpression when compared to neutrophils from type 2 diabetes subjects. Stimulation with TNF-α or LPS increased the expression of ERV-1 by healthy and diabetic neutrophils. RvE1 counteracted LPS and TNF-α induction of ERV-1 overexpression and endogenous diabetic overexpression, activating phagocytosis and resolution signals. Functional ERV-1 was determined by phosphorylation of the signaling protein ribosomal S6. Receptor-antagonism experiments revealed that the increase in phosphorylation of ribosomal S6 was mediated by BLT-1 in healthy subject neutrophils and by ERV-1 in diabetes. Metabololipidomics reveal a proinflammatory profile in diabetic serum. Cell phagocytosis is impaired in type 2 diabetes and requires RvE1 for activation. The dose of RvE1 required to activate resolution signals in type 2 diabetic neutrophils was significantly higher than in healthy controls. RvE1 rescues the dysregulation seen on neutrophil receptor profile and, following a therapeutic dosage, activates phagocytosis and resolution signals in type 2 diabetes. These findings reveal the importance of resolution receptors in health, disease, and dysregulation of inflammation in type 2 diabetes.
Asunto(s)
Reductasas del Citocromo/metabolismo , Diabetes Mellitus Tipo 2/metabolismo , Ácido Eicosapentaenoico/análogos & derivados , Neutrófilos/metabolismo , Receptores de Leucotrieno B4/metabolismo , Adulto , Células Cultivadas , Cromatografía Liquida , Reductasas del Citocromo/inmunología , Diabetes Mellitus Tipo 2/inmunología , Ácido Eicosapentaenoico/inmunología , Ácido Eicosapentaenoico/metabolismo , Femenino , Humanos , Inflamación/inmunología , Inflamación/metabolismo , Masculino , Persona de Mediana Edad , Neutrófilos/inmunología , Oxidorreductasas actuantes sobre Donantes de Grupos Sulfuro , Fagocitosis/inmunología , Reacción en Cadena de la Polimerasa , Receptores de Leucotrieno B4/inmunología , Espectrometría de Masas en Tándem , TranscriptomaRESUMEN
Leukotriene B4 receptor 1 (BLT1), a high-affinity G protein-coupled receptor (GPCR) for leukotriene B4 (LTB4), plays important roles in inflammatory and immune reactions. Although the LTB4-BLT1 axis is known to promote inflammation, the binding proteins that modulate LTB4-BLT1 signaling have not been identified. Recently, we discovered that receptor for advanced glycation end products (RAGE) interacts with BLT1 and modulates LTB4-BLT1 signaling. We propose RAGE as a new class of GPCR modulator and a new target of future GPCR studies.
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Leucocitos Mononucleares/inmunología , Leucotrieno B4/metabolismo , Macrófagos/inmunología , Receptor para Productos Finales de Glicación Avanzada/genética , Receptores de Leucotrieno B4/genética , Regulación de la Expresión Génica , Humanos , Inflamación , Leucocitos Mononucleares/patología , Leucotrieno B4/inmunología , Sistema de Señalización de MAP Quinasas/genética , Sistema de Señalización de MAP Quinasas/inmunología , Macrófagos/patología , Proteína Quinasa 1 Activada por Mitógenos/genética , Proteína Quinasa 1 Activada por Mitógenos/inmunología , Proteína Quinasa 3 Activada por Mitógenos/genética , Proteína Quinasa 3 Activada por Mitógenos/inmunología , FN-kappa B/genética , FN-kappa B/inmunología , Fosforilación , Receptor para Productos Finales de Glicación Avanzada/inmunología , Receptores de Leucotrieno B4/inmunologíaRESUMEN
Bioactive matrix fragments (matrikines) have been identified in a myriad of disorders, but their impact on the evolution of airway inflammation has not been demonstrated. We recently described a pathway where the matrikine and neutrophil chemoattractant proline-glycine-proline (PGP) could be degraded by the enzyme leukotriene A4 hydrolase (LTA4H). LTA4H classically functions in the generation of pro-inflammatory leukotriene B4, thus LTA4H exhibits opposing pro- and anti-inflammatory activities. The physiological significance of this secondary anti-inflammatory activity remains unknown. Here we show, using readily resolving pulmonary inflammation models, that loss of this secondary activity leads to more pronounced and sustained inflammation and illness owing to PGP accumulation. PGP elicits an exacerbated neutrophilic inflammation and protease imbalance that further degrades the extracellular matrix, generating fragments that perpetuate inflammation. This highlights a critical role for the secondary anti-inflammatory activity of LTA4H and thus has consequences for the generation of global LTA4H inhibitors currently being developed.
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Epóxido Hidrolasas/genética , Matriz Extracelular/inmunología , Infecciones por Haemophilus/inmunología , Pulmón/inmunología , Macrófagos Alveolares/inmunología , Neutrófilos/inmunología , Oligopéptidos/inmunología , Neumonía Neumocócica/inmunología , Prolina/análogos & derivados , Animales , Epóxido Hidrolasas/inmunología , Matriz Extracelular/metabolismo , Citometría de Flujo , Haemophilus influenzae tipo b , Inflamación , Elastasa de Leucocito/metabolismo , Leucotrieno B4/inmunología , Metaloproteinasa 12 de la Matriz/metabolismo , Metaloproteinasa 9 de la Matriz/metabolismo , Ratones , Ratones Noqueados , Neumonía Bacteriana/inmunología , Prolina/inmunología , Receptores de Leucotrieno B4/genética , Receptores de Leucotrieno B4/inmunología , Streptococcus pneumoniaeRESUMEN
Leukotriene B4 (LTB4) contributes to many inflammatory diseases, including genetic and nongenetic forms of chronic obstructive pulmonary disease. α-1 Antitrypsin (AAT) deficiency (AATD) is characterized by destruction of lung parenchyma and development of emphysema, caused by low AAT levels and a high neutrophil burden in the airways of affected individuals. In this study we assessed whether AATD is an LTB4-related disease and investigated the ability of serum AAT to control LTB4 signaling in neutrophils. In vitro studies demonstrate that neutrophil elastase is a key player in the LTB4 inflammatory cycle in AATD, causing increased LTB4 production, and associated BLT1 membrane receptor expression. AATD patients homozygous for the Z allele were characterized by increased neutrophil adhesion and degranulation responses to LTB4. We demonstrate that AAT can bind LTB4 and that AAT/LTB4 complex formation modulates BLT1 engagement and downstream signaling events, including 1,4,5-triphosphate production and Ca(2+) flux. Additionally, treatment of ZZ-AATD individuals with AAT augmentation therapy decreased plasma LTB4 concentrations and reduced levels of membrane-bound neutrophil elastase. Collectively, these results provide a mechanism by which AAT augmentation therapy impacts on LTB4 signaling in vivo, and not only reinforces the utility of this therapy for resolving inflammation in AATD, but supports useful future clinical applications in treatment of other LTB4-related diseases.
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Señalización del Calcio/inmunología , Degranulación de la Célula/inmunología , Leucotrieno B4/inmunología , Neutrófilos/inmunología , Receptores de Leucotrieno B4/inmunología , Deficiencia de alfa 1-Antitripsina/inmunología , alfa 1-Antitripsina/inmunología , Adulto , Femenino , Humanos , Elastasa de Leucocito/inmunología , Pulmón/inmunología , Pulmón/patología , Masculino , Neutrófilos/patología , alfa 1-Antitripsina/uso terapéutico , Deficiencia de alfa 1-Antitripsina/tratamiento farmacológico , Deficiencia de alfa 1-Antitripsina/patologíaRESUMEN
Leukotriene B4 (LTB4 ) is a lipid mediator that is rapidly generated in inflammatory sites, and its functional receptor, BLT1, is mostly expressed on immune cells. Contact dermatitis is a common inflammatory skin disease characterized by skin oedema and abundant inflammatory infiltrates, primarily including neutrophils and CD8(+) T cells. The role of the LTB4 -BLT1 axis in contact dermatitis remains largely unknown. In this study, we found up-regulated gene expression of 5-lipoxygenase and leukotriene A4 hydrolase, two critical enzymes for LTB4 synthesis, BLT1 and elevated LTB4 levels in skin lesions of oxazolone (OXA)-induced contact dermatitis. BLT1 deficiency or blockade of LTB4 and BLT1 by the antagonists, bestatin and U-75302, respectively, in the elicitation phase caused significant decreases in ear swelling and skin-infiltrating neutrophils and CD8(+) T cells, which was accompanied by significantly reduced skin expression of CXCL1, CXCL2, interferon-γ and interleukin-1ß. Furthermore, neutrophil depletion during the elicitation phase of OXA-induced contact dermatitis also caused significant decreases in ear swelling and CD8(+) T-cell infiltration accompanied by significantly decreased LTB4 synthesis and gene expression of CXCL2, interferon-γ and interleukin-1ß. Importantly, subcutaneous injection of exogenous LTB4 restored the skin infiltration of CD8(+) T cells in neutrophil-depleted mice following OXA challenge. Collectively, our results demonstrate that the LTB4 -BLT1 axis contributes to OXA-induced contact dermatitis by mediating skin recruitment of neutrophils, which are a major source of LTB4 that sequentially direct CD8(+) T-cell homing to OXA-challenged skin. Hence, LTB4 and BLT1 could be potential therapeutic targets for the treatment of contact dermatitis.
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Linfocitos T CD8-positivos/inmunología , Dermatitis por Contacto/inmunología , Leucotrieno B4/inmunología , Neutrófilos/inmunología , Receptores de Leucotrieno B4/inmunología , Animales , Araquidonato 5-Lipooxigenasa/biosíntesis , Quimiocina CXCL1/biosíntesis , Quimiocina CXCL2/biosíntesis , Dermatitis por Contacto/tratamiento farmacológico , Epóxido Hidrolasas/biosíntesis , Alcoholes Grasos/farmacología , Femenino , Glicoles/farmacología , Inflamación/tratamiento farmacológico , Inflamación/inmunología , Interferón gamma/biosíntesis , Interleucina-1beta/biosíntesis , Leucina/análogos & derivados , Leucina/farmacología , Leucotrieno B4/antagonistas & inhibidores , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Oxazolona , Receptores de Leucotrieno B4/antagonistas & inhibidores , Receptores de Leucotrieno B4/biosíntesis , Piel/citología , Piel/inmunologíaRESUMEN
The potent chemokine leukotriene B4 (LTB4) is increased in obesity, and is associated with insulin resistance. A recent article shows that inhibition of its receptor LTB4 receptor 1 (Ltb4r1) improves insulin sensitivity in muscle and liver via cytokine-independent mechanisms, and reduces proinflammatory immune cell infiltration in the adipose tissue.
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Hepatocitos/inmunología , Resistencia a la Insulina/inmunología , Leucotrieno B4/inmunología , Macrófagos/inmunología , Fibras Musculares Esqueléticas/inmunología , Obesidad/inmunología , Receptores de Leucotrieno B4/inmunología , AnimalesRESUMEN
Insulin resistance results from several pathophysiologic mechanisms, including chronic tissue inflammation and defective insulin signaling. We found that liver, muscle and adipose tissue exhibit higher levels of the chemotactic eicosanoid LTB4 in obese high-fat diet (HFD)-fed mice. Inhibition of the LTB4 receptor Ltb4r1, through either genetic or pharmacologic loss of function, led to an anti-inflammatory phenotype with protection from insulin resistance and hepatic steatosis. In vitro treatment with LTB4 directly enhanced macrophage chemotaxis, stimulated inflammatory pathways, reduced insulin-stimulated glucose uptake in L6 myocytes, and impaired insulin-mediated suppression of hepatic glucose output in primary mouse hepatocytes. This was accompanied by lower insulin-stimulated Akt phosphorylation and higher Irs-1/2 serine phosphorylation, and all of these events were dependent on Gαi and Jnk1, two downstream mediators of Ltb4r1 signaling. These observations elucidate a novel role of the LTB4-Ltb4r1 signaling pathway in hepatocyte and myocyte insulin resistance, and they show that in vivo inhibition of Ltb4r1 leads to robust insulin-sensitizing effects.
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Hepatocitos/inmunología , Resistencia a la Insulina/inmunología , Leucotrieno B4/inmunología , Macrófagos/inmunología , Fibras Musculares Esqueléticas/inmunología , Obesidad/inmunología , Receptores de Leucotrieno B4/inmunología , Animales , Glucemia/metabolismo , Dieta Alta en Grasa , Hígado Graso/inmunología , Hígado Graso/metabolismo , Hepatocitos/metabolismo , Inflamación/inmunología , Insulina/metabolismo , Proteínas Sustrato del Receptor de Insulina/metabolismo , Ratones , Ratones Obesos , Fibras Musculares Esqueléticas/metabolismo , Obesidad/metabolismo , Fosforilación , Proteínas Proto-Oncogénicas c-akt/metabolismo , Receptores de Leucotrieno B4/antagonistas & inhibidores , Receptores de Leucotrieno B4/genética , Transducción de SeñalRESUMEN
Neutrophils play important roles in many inflammatory diseases. The migration of neutrophils to the inflammatory site is tightly regulated by specific chemokines, of which interleukin-8 (IL-8) and leukotriene B4 (LTB4 ) constitute key mediators by binding to the surface receptors CXCR1/2 and BLT1, respectively. Oligonucleotides (ODN) containing CpG motifs mediate potent immunomodulatory effects through binding to Toll-like receptor 9. So far, knowledge on how ODN can affect neutrophil migration during inflammation is lacking. This study demonstrates that several novel CpG ODN significantly down-regulate the surface expression of CXCR1/2 and BLT1. In addition, the ODN significantly blocked IL-8-induced and LTB4 -induced neutrophil migration in vitro, as well as leucocyte migration in vivo demonstrated in mice by intravital microscopy and in a model of airway inflammation. The down-regulation of CXCR1 is rapid, occurring 15 min after ODN stimulation, and can be mediated through an endosomally independent mechanism. Inhibition of the IL-8 and LTB4 pathways may provide new opportunities of therapeutic intervention using ODN to reduce neutrophil infiltration during inflammation.
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Factores Inmunológicos/farmacología , Inmunomodulación , Infiltración Neutrófila/efectos de los fármacos , Neutrófilos/efectos de los fármacos , Oligonucleótidos/farmacología , Receptores de Interleucina-8A/biosíntesis , Receptores de Interleucina-8B/biosíntesis , Receptores de Leucotrieno B4/inmunología , Animales , Quimiotaxis de Leucocito/efectos de los fármacos , Quimiotaxis de Leucocito/inmunología , Islas de CpG/genética , Regulación hacia Abajo/inmunología , Femenino , Humanos , Inflamación/tratamiento farmacológico , Inflamación/inmunología , Interleucina-8/antagonistas & inhibidores , Interleucina-8/biosíntesis , Interleucina-8/inmunología , Antígeno de Macrófago-1/inmunología , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Infiltración Neutrófila/inmunología , Neutrófilos/inmunología , Ovalbúmina , Receptores de Leucotrieno B4/antagonistas & inhibidores , Receptores de Leucotrieno B4/biosíntesis , Receptor Toll-Like 9/inmunología , Factor de Necrosis Tumoral alfa/inmunologíaRESUMEN
Psoriasis is an inflammatory skin disease with accelerated epidermal cell turnover. Neutrophil accumulation in the skin is one of the histological characteristics of psoriasis. However, the precise mechanism and role of neutrophil infiltration remain largely unknown. In this article, we show that orchestrated action of CXCR2 and leukotriene B4 receptor BLT1 plays a key role in neutrophil recruitment during the development of imiquimod (IMQ)-induced psoriatic skin lesions in mice. Depletion of neutrophils with anti-Ly-6G Ab ameliorated the disease severity, along with reduced expression of proinflammatory cytokine IL-1ß in the skin. Furthermore, CXCR2 and BLT1 coordinately promote neutrophil infiltration into the skin during the early phase of IMQ-induced inflammation. In vitro, CXCR2 ligands augment leukotriene B4 production by murine neutrophils, which, in turn, amplifies chemokine-mediated neutrophil chemotaxis via BLT1 in autocrine and/or paracrine manners. In agreement with the increased IL-19 expression in IMQ-treated mouse skin, IL-1ß markedly upregulated expression of acanthosis-inducing cytokine IL-19 in human keratinocytes. We propose that coordination of chemokines, lipids, and cytokines with multiple positive feedback loops might drive the pathogenesis of psoriasis and, possibly, other inflammatory diseases as well. Interference to this positive feedback or its downstream effectors could be targets of novel anti-inflammatory treatment.
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Queratinocitos/metabolismo , Infiltración Neutrófila/inmunología , Psoriasis/inmunología , Receptores de Interleucina-8B/metabolismo , Receptores de Leucotrieno B4/metabolismo , Adyuvantes Inmunológicos/toxicidad , Aminoquinolinas/toxicidad , Animales , Quimiotaxis de Leucocito , Modelos Animales de Enfermedad , Imiquimod , Inmunohistoquímica , Queratinocitos/inmunología , Ratones , Ratones Noqueados , Neutrófilos/inmunología , Psoriasis/inducido químicamente , Psoriasis/metabolismo , Reacción en Cadena en Tiempo Real de la Polimerasa , Receptores de Interleucina-8B/inmunología , Receptores de Leucotrieno B4/inmunologíaRESUMEN
The bioactive lipid mediator leukotriene B4 (LTB4) greatly enhances phagocyte antimicrobial functions against a myriad of pathogens. In murine histoplasmosis, inhibition of the LT-generating enzyme 5-lypoxigenase (5-LO) increases the susceptibility of the host to infection. In this study, we investigated whether murine resistance or susceptibility to Histoplasma capsulatum infection is associated with leukotriene production and an enhancement of in vivo and/or in vitro antimicrobial effector function. We show that susceptible C57BL/6 mice exhibit a higher fungal burden in the lung and spleen, increased mortality, lower expression levels of 5-LO and leukotriene B4 receptor 1 (BLT1) and decreased LTB4 production compared to the resistant 129/Sv mice. Moreover, we demonstrate that endogenous and exogenous LTs are required for the optimal phagocytosis of H. capsulatum by macrophages from both murine strains, although C57BL/6 macrophages are more sensitive to the effects of LTB4 than 129/Sv macrophages. Therefore, our results provide novel evidence that LTB4 production and BLT1 signaling are required for a histoplasmosis-resistant phenotype.
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Histoplasma/inmunología , Histoplasmosis/veterinaria , Leucotrieno B4 , Receptores de Leucotrieno B4/inmunología , Enfermedades de los Roedores/inmunología , Animales , Araquidonato 5-Lipooxigenasa/genética , Araquidonato 5-Lipooxigenasa/inmunología , Susceptibilidad a Enfermedades , Inhibidores Enzimáticos/farmacología , Expresión Génica/inmunología , Histoplasma/patogenicidad , Histoplasmosis/genética , Histoplasmosis/inmunología , Histoplasmosis/metabolismo , Especificidad del Huésped , Interacciones Huésped-Patógeno , Leucotrieno B4/metabolismo , Leucotrieno B4/farmacología , Pulmón/efectos de los fármacos , Pulmón/inmunología , Pulmón/microbiología , Macrófagos/citología , Macrófagos/efectos de los fármacos , Macrófagos/microbiología , Ratones , Ratones de la Cepa 129 , Ratones Endogámicos C57BL , Fagocitosis/efectos de los fármacos , Receptores de Leucotrieno B4/genética , Enfermedades de los Roedores/genética , Enfermedades de los Roedores/metabolismo , Transducción de Señal , Bazo/efectos de los fármacos , Bazo/inmunología , Bazo/microbiologíaRESUMEN
MicroRNAs are known to control TLR activation in phagocytes. We have shown that leukotriene (LT) B4 (LTB4) positively regulates macrophage MyD88 expression by decreasing suppressor of cytokine signaling-1 (SOCS-1) mRNA stability. In this study, we investigated the possibility that LTB4 control of MyD88 expression involves the generation of microRNAs. Our data show that LTB4, via its receptor B leukotriene receptor 1 (BLT1) and Gαi signaling, increased macrophage expression of inflammatory microRNAs, including miR-155, miR-146b, and miR-125b. LTB4-mediated miR-155 generation was attributable to activating protein-1 activation. Furthermore, macrophage transfection with antagomirs against miR-155 and miR-146b prevented both the LTB4-mediated decrease in SOCS-1 and increase in MyD88. Transfection with miR-155 and miR-146b mimics decreased SOCS-1 levels, increased MyD88 expression, and restored TLR4 responsiveness in both wild type and LT-deficient macrophages. To our knowledge, our data unveil a heretofore unrecognized role for the GPCR BLT1 in controlling expression of microRNAs that regulate MyD88-dependent activation of macrophages.